Characterization of pulmonary progenitor cell dysfunction in COPA syndrome

项目摘要-摘要 Coatomer蛋白α亚基（COPA）综合征是一种罕见的常染色体显性单基因疾病， 导致影响肺部和关节的多器官疾病，并伴有不完全的呼吸道感染。患者出现早 在生活中患有间质性肺病（ILD），并具有升高的炎症标志物和阳性自身抗体滴度。 虽然存在控制这种异常免疫表型的治疗方法，但它们不能解决未知的问题。 这种疾病的机制。COP β是一种广泛表达的外壳体蛋白复合物I亚基 （COPI），其负责蛋白质从高尔基体到ER的逆行运输。改动或 该途径的中断导致细胞应激。到目前为止，大多数研究都研究了免疫系统的作用。 系统在COPA综合征的发病和进展中的作用，而没有阐明这种疾病的原因。因此，我们认为， 我们相信还涉及其他细胞类型和过程。大多数COPA患者在某个时候会患上 肺部疾病谱，如细支气管炎、ILD和/或肺纤维化（PF）。这些疾病共享 某些致病特征响应于细胞应激，包括但不限于ER应激， 未折叠蛋白反应（UPR）活化、活化的成纤维细胞、炎症和异常自噬。 近端和远端肺的祖细胞、基底和肺泡II型细胞（ATII）分别是 受炎症影响，导致整个气道上皮细胞的异常维持。我们认为 在COPA综合征患者中发生祖细胞功能障碍的类似机制。因此我们 假设COPA综合征是由一种内在的祖细胞缺陷引起的， 上皮维持导致ILD和肺纤维化。在目标1中，我们将确定上皮细胞- 由COPAMT表达引起的固有缺陷。这将通过将缔约方会议的特点 相互作用组来阐明导致细胞应激的异常蛋白质相互作用。然后我们将描述 使用单细胞RNA测序突出差异基因表达模式的COPAMT转录组 与疾病有关。此外，我们将利用体内小鼠模型来准确地描述这种联系。 肺祖细胞功能障碍与COPA相关免疫反应激活之间的关系 综合征在目标2中，我们将使用以下方法验证挽救COPA综合征上皮细胞缺陷的策略： 慢病毒载体递送系统和基因编辑工具，目的是挽救患病的表型。

PROJECT SUMMARY – ABSTRACT Coatomer protein subunit alpha (COPA) syndrome is a rare, autosomal dominant, monogenic disorder that results in a multiorgan disease affecting the lungs and joints with incomplete penetrance. Patients present early in life with interstitial lung disease (ILD) and have elevated inflammatory markers and positive autoantibody titers. While treatments exist to control this aberrant immune phenotype, they do not address the unknown mechanism(s) of this disease. COP is a ubiquitously expressed subunit of the coatomer protein complex I (COPI) which is responsible for retrograde transport of proteins from the Golgi to the ER. Alterations or disruptions in this pathway result in cellular stress. To date, most studies have examined the role of the immune system in the onset and progression of COPA syndrome without elucidating a cause for this disease. Therefore, we believe other cell types and processes are involved. Most COPA patients will at some point suffer from a spectrum of pulmonary disease such as bronchiolitis, ILD, and/or pulmonary fibrosis (PF). These diseases share certain pathogenic characteristics in response to cellular stress including, but not limited to, ER stress and unfolded protein response (UPR) activation, activated fibroblasts, inflammation, and aberrant autophagy. Progenitor cells of the proximal and distal lung, basal and alveolar type II cells (ATII), respectively, are disproportionally affected causing aberrant maintenance of the epithelium throughout the airway. We believe similar mechanisms of progenitor cell dysfunction are occurring in COPA syndrome patients. We therefore hypothesize that COPA syndrome is driven by an intrinsic progenitor cell defect that compromises epithelial maintenance resulting in ILD and lung fibrosis. In Aim 1, we will determine the epithelial cell- intrinsic defects resulting from the expression of COPAMT. This will be achieved by characterizing the COP interactome to elucidate anomalous protein interactions leading to cellular stress. We will then characterize the transcriptome of COPAMT using single cell RNA sequencing to highlight differential gene expression patters associated with disease. Furthermore, we will utilize in vivo mouse models to accurately describe the link between pulmonary progenitor cell dysfunction and activation of the immune responses associated with COPA syndrome. In Aim 2, we will validate strategies to rescue the epithelial cell defect of COPA syndrome using lentiviral vector delivery systems and gene editing tools with the intention of rescuing the diseased phenotype.